Types of Media for Microbiology

Blood Agar A rich, non-selective, differential medium which supports the growth of a
wide variety of microbes. It is made by adding 5% sheeps blood to a complex medium
containing peptones just before the plates are poured. The red blood cells remain intact
and the medium resembles fresh blood. Some bacteria grow on blood agar without
disturbing the red cells; others hemolyze the cells to various degrees. Bacteria that lyse
the red cells with a complete clearing of the medium around the colonies are referred to
as beta-hemolytic. Some bacteria lyse the cells but do not break down the hemoglobin
completely leaving greenish methemoglobin in the medium. This is called alphahemolysis. Non-hemolytic bacteria have no visible effect on the red blood cells and are
said to be gamma-hemolytic.
Eosin Methylene Blue Agar (EMB) This agar medium is both selective and differential.
The combination of the two dyes eosin and methylene blue inhibits most Gram positive
bacteria but allows many Gram negative organisms to grow. In addition to peptones,
EMB contains lactose (it may also contain sucrose). Gram negative bacteria that
ferment the lactose produce acid which turns the colonies dark purple as the acid acts
upon the dyes. In addition, certain lactose-fermenting bacteria produce flat, dark
colonies with a green metallic sheen. Other lactose fermenters produce larger, mucoid
colonies, often purple only in their center. Lactose non-fermenters are either colorless or
light lavender.
MacConkey Agar (MAC) This is a differential plating medium, selective for Gram
negative organisms, used primarily for detection and isolation of enteric bacteria. Grampositives are generally inhibited by crystal violet in the medium. Isolated colonies of
lactose-fermenting bacteria are brick red in color and may be surrounded by a zone of
precipitated bile. This reaction is due to the action of the acids, produced by
fermentation of lactose, upon bile salts present in the medium, and subsequent
absorption of neutral red. Non-lactose fermenters are colorless and transparent.
Mannitol Salt Agar (MSA) This is a selective medium that favors the growth of
staphylococci and for differentiation between pathogenic and non-pathogenic types.
MSA contains 7.5% NaCl and staphylococci are characteristically able to tolerate this
high salt concentration. MSA also contains the sugar-alcohol mannitol and the pH
indicator phenol red. Generally pathogenic staphylococci are able to ferment the
mannitol, lowering the pH, and thereby turning the indicator yellow The non-pathogenic
staphylococci do not ferment mannitol and the medium remains pink in their vicinity.
Milk Agar This is nutrient agar containing powdered mild. It is used to test for an
organisms ability to secrete extracellular proteases that catalyze the hydrolysis of the
milk protein casein. A clear zone around a colony indicates casein hydrolysis.
Phenylethanol Agar (PEA) Phenylethanol inhibits many Gram negative bacteria and
its inclusion in PEA renders the medium selective for Gram positives.

Sabouraud Dextrose Agar (SDA) A medium that promotes the growth of fungi (yeast
and molds) but which is inhibitory for many bacteria
Simmons Citrate Agar This medium, which may be used in the form of slants or
plates, tests for the ability of an organism to grow on citrate as the sole carbon source. It
contains various minerals, including ammonium phosphate for nitrogen, sodium citrate,
agar, and the pH indicator brom thymol blue. The indicator is green when acid or
neutral. As bacteria grow on the medium, they make the medium increasingly alkaline
and the indicator turns blue.
Spirit Blue Agar This medium is used for detecting lipase activity. It contains nutrients,
emulsified lipids and the dye spirit blue. Clearing of the emulsion is a positive indicator
of lipase. Some organisms will bring about precipitation of the blue dye.
Trypticase Soy Agar (TSA) A general purpose nutrient medium that promotes the
growth of bacteria